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bioRxiv · 10.1101/2021.03.19.436169

Visualising formation of the ribosomal active site in mitochondria

Chandrasekaran, V.·Desai, N.·Burton, N. O.·Yang, H.·Price, J. L.·Miska, E. A.·Ramakrishnan, V.
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Abstract

Ribosome assembly is an essential and complex process that is regulated at each step by specific biogenesis factors. Using cryo-electron microscopy, we identify and order major steps in the formation of the highly conserved peptidyl transferase centre (PTC) and tRNA binding sites in the large subunit of the human mitochondrial ribosome (mitoribosome). The conserved GTPase GTPBP7 regulates the folding and incorporation of core 16S ribosomal RNA (rRNA) helices and the ribosomal protein bL36m, and ensures that the PTC base U3039 has been 2'-O-methylated. Additionally, GTPBP7 binds the RNA methyltransferase NSUN4 and MTERF4, which facilitate earlier steps by sequestering H68-71 of the 16S rRNA and allowing biogenesis factors to access the maturing PTC. Consistent with the central role of NSUN4*MTERF4 and GTPBP7 during mitoribosome biogenesis, in vivo mutagenesis designed to disrupt binding of their Caenorhabditis elegans orthologs to the large subunit potently activates mitochondrial stress responses and results in severely reduced viability, developmental delays and sterility. Next-generation RNA sequencing reveals widespread gene expression changes in these mutant animals that are indicative of mitochondrial stress response activation. We also answer the long-standing question of why NSUN4 but not its enzymatic activity, is indispensable for mitochondrial protein synthesis in metazoans.

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Chandrasekaran, V., Desai, N., Burton, N. O., Yang, H., Price, J. L., Miska, E. A., Ramakrishnan, V.. 2021-03-20. Visualising formation of the ribosomal active site in mitochondria. https://doi.org/10.1101/2021.03.19.436169

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